英文摘要

Question

摘要
目的: 用基因工程技术构建含pGEX-4T-1-has2基因的重组质粒, 获得Has2活性蛋白,制备抗体的探针。
方法：首先,上网获得基因序列，运用premier5.0软件设计引物，运用巢式PCR技术扩增has2基因，第一对引物做PCR扩增后，电泳检测结果是正确的500bp至600bp之间后，再以PCR产物为模板,以带酶切位点的第二对引物做PCR扩增，电泳检测正确后，跑纯化胶，进行纯化回收，将纯化回收产物连pMD18-T进行转化，然后挑单克隆以及摇菌，小提质粒，再进行双酶切检测和PCR检测，当两项结果均正确时，送pMD18-T—has2新鲜菌液去测序。测序正确后，大体系双酶切pMD18-T- has2，纯化回收后将has2连接入pGEX4T-1载体，同样进行转化、挑单克隆、摇菌、提质粒，得到pGEX-4T-1—has2重组质粒。
结果:获得重组质粒，酶切及测序结果正确。
结论: pGEX-4T-1—has2重组质粒构建成功. pGEX-4T-1—has重组质粒的构建及鉴定结果正确

Answer 1

Abstract
Objective: To use genetic engineering technology building with pGEX-4T-1-has2 the recombinant gene was Has2 activity of proteins, antibody preparation of the probe.
Method: First of all, Internet access to genetic sequence, use premier5.0 software design primer, use nested PCR amplification has2 gene, the first primer to do PCR amplification, electrophoresis test results are correct 500 bp to 600 bp between after And then to PCR product as a template to bring restriction site of the second pair of primers do PCR amplification, electrophoresis test correctly, ran purification plastic, purification recovery, recycling and purification products for even the pMD18-T Conversion, and then pick monoclonal and shake of small to plasmid and then double-digested PCR testing and detection, when the two results are correct, sending pMD18-T-has2 fresh bacilli to sequencing. After the correct sequence, the system of dual-digestion pMD18-T-has2, purification recovery after has2 connected to the pGEX4T-1 carrier, the same conversion, pick monoclonal, shake bacteria, to plasmid be pGEX-4T-1 - Has2 recombinant.
Results: recombinant, digestion and the correct sequence.
Conclusion: pGEX-4T-1-has2 recombinant Construction of success. PGEX-4T-1-has recombinant Construction and identification of the correct results